目的：探讨不同浓度胆红素对新生儿脐血单核细胞（CBMC）表面分子Toll样受体4（TLR4）的影响。方法：采用纤维连接蛋白结合法分离得到CBMC，将其加入含不同浓度胆红素（0~307.8 μmol/L）的牛血清白蛋白溶液中孵育1 h，再加入脂多糖（LPS，1 μg/mL）刺激24 h。收集CBMC，采用间接免疫荧光技术检测细胞表面分子TLR4的表达。结果：胆红素浓度为102.6、153.9、 220.6和307.8 μmol/L时，CBMC表面分子TLR4 的表达率均明显降低，随着胆红素浓度的升高，这种降低趋势越明显。结论：胆红素抑制CBMC表面分子TLR4的表达，随着胆红素浓度的升高，抑制作用越明显。
Abstract
OBJECTIVE: To study the effect of different concentrations of bilirubin on expression of toll-like receptor 4 (TLR4) in cord blood monocytes (CBMC). METHODS: Under the sterile condition, umbilical vein blood samples were obtained from normal full-term newborns, and the monocytes were in vitro separated by the method of gelatin/plasma coated flasks. The monocytes were preincubated with various concentrations (0-307.8 μmol/L) of bilirubin dissolved in bovine albumin solution for 1 hr. Bilirubin-treated CBMC were further cultured with LPS (1 μg/mL) to induce cellular activation for 24 hrs, and then the CBMC were collected. The expression of TLR4 in monocytes was measured by indirect immunofluorescence method. RESULTS: Bilirubin at the concentrations of 102.6, 153.9, 220.6 and 307.8 μmol/L inhibited the expression of TLR4 of CBMC. The inhibition effect increased with the increasing concentration of bilirubin. CONCLUSIONS: Bilirubin can inhibit the TLR4 expression of CBNC in a dose-dependent manner.
关键词
胆红素 /
Toll样受体4 /
脐血 /
单核细胞
Key words
Bilirubin /
Toll-like receptor 4 /
Cord blood /
Monocyte
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参考文献
［1］ Liu Y, Li P, Lu J, Xiong W, Oger J, Tetzlaff W, et al. Bilirubin possesses powerful immuno-modulatory activity and suppresses experimental auto-immune encephalomyelitis［J］. J Immunol, 2008, 181(3): 1887-1897.
［2］林丽星，余唯琪，张晓燕，李玉梅，陶仲宾.高胆红素血症新生儿红细胞免疫功能的变化及其影响因素［J］.实用儿科临床杂志，2007，22（6）：414-415.
［3］张宇，孙瑞利，胡锦跃.Toll样受体4信号转导研究进展［J］.国际病理科学与临床杂志，2009，29（1）：32-36.
［4］裴雪梅，陈昌辉，牛会琴，吴青，李茂军.不同浓度胆红素对新生儿脐血单核细胞形态结构的影响［J］.医学研究生学报，2009，22（5）：496-499.
［5］牛会琴，陈昌辉，裴雪梅，吴青，李茂军. 胆红素对新生儿脐血单核细胞吞噬功能的影响［J］.中国新生儿科杂志，2009，24（2）：115-117.
［6］裴雪梅，陈昌辉，牛会琴，吴青，李茂军.不同浓度胆红素对新生儿脐血单核细胞白细胞介素-6、肿瘤坏死因子α分泌的影响［J］.实用儿科临床杂志，2009，24（2）：102-103.
［7］Freundlich B, Avdalovic N. Use of gelatin/plasma coated flasks for isolating human peripheral blood monocytes［J］. Immunol Methods, 1983, 62（1）：31-37.
［8］江志雄，金益人，洪文澜，尚世强，孙眉月.新生儿脐血单核细胞在脂多糖刺激下分泌IL-6及出现IL-6mRNA表达的实验研究［J］.中国免疫学杂志，2002，18:862-865.
［9］何凤莲，张妍蓓.Toll样受体的研究进展［J］.国际呼吸杂志，2006，26 (9):655-659.
［10］Daly KA, Lefevre C, Nicholas K, Deane E, Williamson P. CD14 and TLR4 are expressed early in tammar (Macropus eugenii) neonate development［J］. J Exp Biol, 2008, 211（8）: 1344-1351.
［11］Forster Waldl E, Sadeghi K, Tamandl D, Gerhold B, Hallwirth U, Rohrmeister K, et al. Monocyte toll-like receptor 4 expression and LPS-induced cytokine production increase during gestational aging［J］.Pediatr Res, 2005, 58(1): 121-124.
［12］Yan SR, Qing G, Byers DM, Stadnyk AW, Alhertani W, Bortolussi R, et al. Role of MyD88 in diminished tumor necrosis factor alpha production by newborn mononuclear cells in response to lipopolysaccharide［J］. Infect Immun, 2004, 72(3): 1223-1229.
［13］王琳，徐建波，田元，刘亚兰，吴河水.新生儿脐血Toll样受体变化及其意义［J］.中华儿科杂志，2007，45（50）：365-368.
［14］Marodi L. Neonatal innate immunity to infectious agents［J］. Infect Immun, 2006, 74(4): 1999-2006.
［15］Wynn JL, Scumpia PO, Winfield RD, Delano MJ, Kelly-Scumpia K, Barker T, et al. Defective innate immunity predisposes murine neonates to poor sepsis outcome but is reversed by TLR agonists［J］.Blood, 2008, 112（5）: 1750-1758.
［16］LeBouder E, Rey-Nores JE, Raby AC, Affolter M, Vidal K, Thornton CA, et al. Modulation of neonatal microbial recognition: TLR-mediated innate immune responses are specifically and differentially modulated by human milk［J］. J Immunol, 2006, 176(6): 3742-3752.
［17］Watchko JF. Hyperbilirubinemia and bilirubin toxicity in the late preterm infant［J］. Clin Perinatol, 2006, 33(4): 839-852.
［18］Stocker R, Yamamoto Y, McDonagh AF, Glazer AN, Ames BN. Bilirubin is an antioxidant of possible physiological importance［J］. Science, 1987, 235(4792): 1043-1046.
［19］Miler I, Sima P, Vetvicka V, Indrova M, Slavikova M. The potential immunosuppressive effect of bilirubin［J］. Allerg Immunol (Leipz), 1988, 34(3): 177-184.
［20］Brito MA, Silva RF, Brites D. Bilirubin toxicity to human erythrocytes: a review［J］.Clin Chim Acta, 2006, 374(1-2): 4656.
［21］Ostrow JD, Tiribelli C. Bilirubin, a curse and a boon［J］. Gut, 2003, 52(12): 1668-1670.
［22］Falcao AS, Silva RF, Pancadas S, Fernandes A, Brito MA, Brites D. Apoptosis and impairment of neurite network by short exposure of immature rat cortical neurons to unconjugated bilirubin increase with cell differentiation and are additionally enhanced by an inflammatory stimulus［J］. J Neurosci Res, 2007, 85(6): 1229-1239.
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